When grown on hydrocarbons, P. oleovorans produces a non-heme iron containing enzyme that hydroxylates the terminal methyl groups of alkanes and fatty acids. We are investigating the action of this omega-hydroxylase on substituted alkanes and fatty acids. Some of these compounds are designed to provide information about the hydroxylation mechanism; others are potential suicide inactivators of the hydroxylase. It is hoped that any inactivators that are found will be prototypes for inactivators of other hydroxylases. A major portion of the work will involve investigating the action of the enzyme on compounds bearing a terminal cyclopropylmethyl group and on compounds bearing a halogen atom on the penultimate carbon. We will look carefully for rearranged products that will provide evidence for intermediates at the carbon undergoing hydroxylation. We will also look for enzyme inactivation that might be traceable to rearrangement or fragmentation of a reactive intermediate. If inactivation is observed it will be studied by kinetic and radiochemical techniques. We eventually plan to extend our work to include a cytochrome P-450 omega-hydroxylase.